The relationship between species within the Xiphinema americanum-group (Nematoda: Dorylaimida)1

Principal components and hierarchical cluster analysis based on morphometric characters of the type populations of the 39 species attributed to the Xiphinema americanum-group confirmed the occurrence of five subgroups. The use of a single character is generally insufficient to characterize a species because of the wide intra-population and intra-specific variations. Hierarchical cluster analysis, although impractical for species/population identification, appears useful in the study of the relationships between species or groups of species.     

A new species of the genus Xiphinema Cobb 1913 (Nematoda: Longidoridae) from north and northwest Iran

European Journal of Plant Pathology - Five populations of a new dagger nematode species were recovered from natural grasslands and forests of north and northwest Iran, and described based upon...     

Correction to: A new species of the genus Xiphinema Cobb 1913 (Nematoda: Longidoridae) from north and Northwest Iran

European Journal of Plant Pathology - A Correction to this paper has been published: https://doi.org/ https://doi.org/10.1007/s10658-021-02272-1     

Molecular characterisation and diagnostics of some Longidorus species (Nematoda: Longidoridae) from Russia and other countries using rRNA genes

The needle nematodes of the genus Longidorus can cause diseases of various crops and trees, and are comprised of more than 150 valid species. Eleven valid and six unidentified species of the genus Longidorus collected in different regions of Russia, two states of USA, Germany, New Zealand and Ukraine were molecularly characterized using analysis of the partial 18S rRNA and the D2–D3 expansion segments of the 28S rRNA gene sequences. Fifty-four partial 28S rRNA and fifteen partial 18S rRNA gene sequences were obtained for the present study. Using molecular criteria, we confirmed the morphological identification and distinguished between the following species: L. aetnaeus, L. africanus, L. andalusicus, L. artemisiae, L. caespiticola, L. distinctus, L. elongatus, L. euonymus, L. intermedius, L. leptocephalus and L. lignosus. Two longidorid populations from Russia and four from California were not identified to a species level. We obtained the full length D2–D3 of 28S rRNA gene sequence from several freshly-collected L. artemisiae samples. We confirmed the identity of the D2 region of 28S rRNA gene sequence with a short D2 of 28S rRNA gene fragment sequence previously obtained from formalin-fixed nematodes embedded in the L. artemisiae paratype slides. Longidorus lignosus was molecularly characterized and L. aetnaeus was reported from Russia for the first time. PCR-D2-D3-RFLP diagnostic profiles generated by five restriction enzymes: AluI, HinfI, Bsp143I, Tru1I and RsaI are presented for sixteen Longidorus species.     

Population structure and species delimitation of rice white tip nematode,Aphelenchoides besseyi (Nematoda: Aphelenchoididae), in China

Aphelenchoides besseyi is a major nematode pathogen in rice known as the causal agent of white tip disease. In the present study, A. besseyi was recovered from 32 seed samples collected from the major rice-growing regions in China. Nine mitochondrial COI haplotypes and 95 ITS genotypes were identified, suggesting a high genetic diversity and endemism level of A. besseyi in China. Therefore, A. besseyi is likely to have experienced a long history of host–parasite coevolution in China, rather than being recently introduced from other countries. The results of population structure analysis indicated the presence of four clusters within Chinese A. besseyi, but these did not correspond to their geographic distribution. Haplotype H2 was found to be the most widespread. Haplotypes H9 and H13 were most genetically divergent and have been recognized as two cryptic species by molecular species delimitation methods, and with limited support from morphometric measurements. The COI-based phylogeny suggested the ability to parasitize rice has independently evolved at least four times in the genus Aphelenchoides.     

Phylogenetic relationships of some spirurine nematodes (Nematoda: Chromadorea: Rhabditida: Spirurina) parasitic in fishes inferred from SSU rRNA gene sequences

Abstract: Small subunit rRNA sequences were obtained from 38 representatives mainly of the nematode orders Spirurida (Camallanidae, Cystidicolidae, Daniconematidae, Philometridae, Physalopteridae, Rhabdochonidae, Skrjabillanidae) and, in part, Ascaridida (Anisakidae, Cucullanidae, Quimperiidae). The examined nematodes are predominantly parasites of fishes. Their analyses provided well-supported trees allowing the study ofphylogenetic relationships among some spirurine nematodes. The present results support the placement of Cucullanidae at the base of the suborder Spirurina and, based on the position of the genus Philonema (subfamily Philoneminae) forming a sister group to Skrjabillanidae (thus Philoneminae should be elevated to Philonemidae), the paraphyly of the Philometridae. Comparison of a large number of sequences of representatives of the latter family supports the paraphyly of the genera Philometra, Philometroides and Dentiphilometra. The validity of the newly included genera Afrophilometra and Caranginema is not supported. These results indicate geographical isolation has not been the cause of speciation in this parasite group and no coevolution with fish hosts is apparent. On the contrary, the group of South-American species ofAlinema, Nilonema and Rumai is placed in an independent branch, thus markedly separated from other family members. Molecular data indicate that the skrjabillanid subfamily Esocineminae (represented by Esocinema bohemicum) should be either elevated to the rank of an independent family or Daniconematidae (Mexiconema africanum) should be decreased to Daniconematinae and transferred to the family Skrjabillanidae. Camallanid genera Camallanus and Procamallanus, as well as the subgenera Procamallanus and Spirocamallanus are confirmed to be paraphyletic. Paraphyly has also been found within Filarioidea, Habronematoidea and Thelazioidea and in Cystidicolidae, Physalopteridae and Thelaziidae. The results of the analyses also show that Neoascarophis, Spinitectus and Rhabdochona are monophyletic, in contrast to the paraphyletic genus Ascarophis. They further confirm the independence of two subgenera, Rhabdochona and Globochona, in the genus Rhabdochona. The necessity of further studies of fish-parasitizing representatives of additional nematode families not yet studied by molecular methods, such as Guyanemidae, Lucionematidae or Tetanonematidae, is underscored.     

Molecular characterization of <Emphasis Type="Italic">Xiphinema brevicollum</Emphasis> (Nematoda: Longidoridae) from the Czech Republic

Populations of Xiphinema brevicollum occurring in the Czech Republic were described morphologically and molecularly. Published species-specific primer set BL18 and BV3 was used to amplify three populations of X. brevicollum from the Czech Republic. These primers were tested against 9 species of Xiphinema and 11 species of Longidorus. Amplification was also observed for X. inaequale, X. italiae and X. lambertii. Three additional markers, mitochondrial cytochrome c oxidase subunit 1, ribosomal D2/D3 expansion segment of the 28S gene and 18S gene, were amplified and sequenced for X. brevicollum, X. inaequale and X. lambertii belonging to X. americanum-group. Comparison of cox1 sequences of X. brevicollum from the Czech Republic with X. taylori from the Slovak Republic (accession number AM086702) suggested that these populations represent the same species.     

Description of Longidorus armeniacae n. sp. (Nematoda: Longidoridae), associated with Prunus armeniaca L. in Semnan province,Iran

European Journal of Plant Pathology - Longidorus armeniacae n. sp. is described and illustrated using morphological and molecular data. It was recovered from the rhizosphere of apricot trees in...     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.     

Morphological and molecular characterisation of Trichodorus golestanensis (Nematoda: Trichodoridae), a new species from Iran

Several species of trichodorid nematodes cause economically important disease on plants. Trichodorus comprises most of the trichodorid species. Trichodorus golestanensis n. sp. is described from a forest park in northern Iran. The new species belongs to the T. lusitanicus morpho-species group and is characterized in male by three ventromedian cervical papillae of which two at level of onchiostyle region, three ventromedian precloacal supplements, the posterior one just anterior to retracted spicules and ventrally curved spicules with mid-blade indentation provided with a few bristles. Females are distinguished by well developed triangular vaginal sclerotized pieces in lateral optical view, a rhomboid-shaped vagina and a transverse slit-like vulva in ventral view. Molecular analysis using the sequence of D2-D3 expansion segment of 28S rDNA differentiated the new species and confirmed the relationships of T. golestanensis n. sp. with T. andalusicus and T. asturanus. The sequences of D2-D3 regions were also provided for previous trichodorid records from Iran T. gilanensis, P. teres and N. minor together with discussion of their relationship.     

Genetic diversity of the weed species,Stellera chamaejasme,in China inferred from amplified fragment length polymorphism analysis

Stellera chamaejasme is a perennial weed with a wide geographic range that is found from the Altai of eastern Russia, northern China and Mongolia southwards as far as the western Himalayas of the Qinghai–Tibet and Yungui Plateaus. The genetic diversity and population structure of 17 populations of S. chamaejasme, represented by 349 individuals, were assessed by using amplified fragment length polymorphism markers. The results showed a relatively high level of genetic variation at the species level. The proportion of total diversity among populations was 0.4370, suggesting significant genetic differentiation and a low gene flow among the populations of this species. The Mantel test indicated that genetic differentiation among populations was significantly correlated with geographic distance. Genetic drift through range expansion and a low gene flow among populations might result in a lower diversity in peripheral populations, compared to central populations. A Bayesian analysis revealed two potential gene pools in S. chamaejasme, which was confirmed by neighbor‐joining clustering and principal coordinate analysis. These results demonstrate that it is necessary to develop suitable biocontrol agents for populations with different gene pools.     

Structure of the cephalic end of Ascarophis mexicana (Nematoda: Cystidicolidae), as revealed by SEM

Scanning electron microscopy of the cystidicolid nematode Ascarophis mexicana Moravec, Salgado-Maldonado et Vivas-Rodríguez, 1995 enabled the first detailed study of its cephalic structure. In contrast to most Ascarophis species, its pseudolabia are highly reduced and sublabia are unlobed and weakly developed. Similascarophis is considered a synonym of Ascarophis, to which two its species are transferred as A. maulensis (Mu?oz, González et George-Nascimento, 2004) comb. n. and A. chilensis (Mu?oz, González et George-Nascimento, 2004) comb. n.     

A new species of Litomosoides (Nematoda: Onchocercidae), parasite of Nectomys palmipes (Rodentia: Cricetidae: Sigmodontinae) from Venezuela: description, molecular evidence, Wolbachia pipientis screening

Abstract: The onchocercid filaria Litomosoides taylori sp. n. is described from the sigmodontine cricetid Nectomys palmipes Allen et Chapman in northeast Venezuela. A voucher specimen of the new species was used for molecular analysis of the coxI and 12S rDNA genes, and screened for the presence of the endobacterium Wolbachia pipientis. Litomosoides taylori belongs to the "sigmodontis group" of Litomosoides and a combination of characters can be used to distinguish it from the remaining 18 species forming this group. Among the five Nectomys species, all living near running water, N. squamipes also harbours Litomosoides species, L. khonae in Brazil and L. navonae in Argentina. These three Litomosoides species of the "sigmodontis group" do not share any particular characters. Gene sequences of L. taylori differ from those of the five Litomosoides species available, the three of the "carinii group" being the most distant. The new species harbours W pipientis, which is concurrent with the great majority of Litomosoides species screened to date.     

Frequency of transmission of artichoke Italian latent nepovirus by Longidorus fasciatus (Nematoda: Longidoridae) from artichoke fields in the Iria and Kandia areas of Argolis in northeast Peloponnesus, Greece

Artichoke Italian latent nepovirus (AILV) transmitted by Longidorus fasciatus is a causative agent of artichoke patchy chlorotic stunting (APCS) in northeast Peloponnesus in southern Greece. Populations of L. fasciatus collected from the Iria and Kandia areas of Argolis in northeast Peloponnesus were used in laboratory experiments to determine the frequency of transmission of the virus by its natural vector. One tenth to almost one half of the L. fasciatus specimens recovered from soil collected in two artichoke fields showing APCS transmitted AILV. Allowing nematodes access for 4 wk to Nicotiana clevelandii mechanically infected with AILV did not increase the number of individual specimens able to transmit virus. The total number of specimens transmitting virus in an experiment did not exceed fifty percent of the individuals tested. Virus-like particles were only observed adsorbed to the inner surface of the odontostyle and it is suggested that the high frequency of transmission of AILV by L. fasciatus is a result of efficient dissociation of virus particles from the specific sites of retention in the vector.     

Rhabdochona minima sp. n. (Nematoda: Spiruroidea) from the loach, Noemacheilus inglisi (Hora), of Nepal.

A new nematode species, Rhabdochona minima sp.n., is described from the intestine of Noemacheilus inglisi (HORA) OF Nepal (River Ganges drainage system). Characteristics features of this species (only males were found) are the small size of body, size and type of spicules, shape of tail and arrangement of caudal papillae. On the basis of its morphology, R. minima sp.n. belongs to the subgenus Globochona sensu Moravec, 1975. The species Rhabdochona cavasius Rehana et Bilqees, 1973 is considered to be a synonym of R. mazedi Prassad et Sahay, 1965.     

Use of ITS nuclear sequences from Phelipanche aegyptiaca as a direct tool to detect single seeds of broomrape species in the soil

Broomrape (Phelipanche and Orobanche spp.) are obligate holoparasites that attack roots of almost all economically-important crops in semiarid regions of the world. Broomrape seeds are extremely small (dust-like seeds), averaging 200 to 300?μm in size and because of the miniscule seed size it is difficult to detect and confirm via conventional methods. In this study our aim was to develop a PCR-based assay specific for broomrape soil-borne seeds and sensitive enough to detect a single or few broomrape seeds in a soil sample. For this purpose, we used complementary polymerase chain reaction (PCR) primers based upon unique sequences in the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA of Phelipanche aegyptiaca. Genomic DNA was extracted from soil samples artificially infested with broomrape seeds or tissue of Phelipanche aegyptiaca Pers., Orobanche cumana Wallr. and Phelipanche crenata Forsk. and subjected to PCR analysis. Using ITS-350 primers, a specific PCR product (350?bp) was amplified and detected in all samples containing broomrape species, but was not detected in soil sample free of broomrape seeds or tissues. Additionally, the PCR-based assay was sensitive enough to detect even a single broomrape seed in the soil. As expected the universal internal control primers amplified a PCR product (555?bp) of genomic DNA extracted from soil samples with or without broomrape tissues or seeds. This diagnostic method is simple, reliable and rapid and could help for assessment of broomrape seed contamination in a crop field.     

Molecular variability and phylogenetic relationships among different species and populations of <Emphasis Type="Italic">Pratylenchus</Emphasis> (Nematoda: Pratylenchidae) as inferred from the analysis of the ITS rDNA

Sequence comparisons and molecular phylogenetic analyses were used to describe the nucleotide variability of the ITS containing regions of eighteen Pratylenchus species and several populations. Comparative analysis of nucleotide sequences of the rDNA internal transcribed spacers (ITS1 and ITS2) among Pratylenchus species used in the present study demonstrates that ITS sequences can widely vary in primary sequence and length. Alignment of eighty-seven Pratylenchus sequences and one outgroup taxon reveals the presence of ambiguous regions that have the greatest effect on phylogeny reconstruction. Phylogenetic analyses using Bayesian Inference, Neighbour Joining-LogDet, Maximum Likelihood and Maximum Parsimony, distinguished twelve highly or moderately supported major clades within Pratylenchus. Our results support the taxonomic usefulness of the ITS region to identify root-lesion nematode species of the genus Pratylenchus but the high nucleotide variability, sometimes, can preclude its use to resolve relationships among all members of the genus. In addition, the phylogenetic groupings are not congruent with those defined by characters derived by lip patterns and numbers of lip annuli.     

Taxonomical study into two new species of Philometra (Nematoda: Philometridae) previously identified as Philometra lateolabracis (Yamaguti, 1935)

Males of the nematode Philometra lateolabracis (Yamaguti, 1935), the type species of the genus Philometra Costa, 1845, were discovered for the first time in gonads of its type host, the Japanese seaperch, Lateolabraxjaponicus (Cuvier). Morphological comparisons carried out between the collected male and female P. lateolabracis with the male and female philometrid nematodes previously reported as P. lateolabracis infecting chicken grunt, Parapristipoma trilineatum (Thunberg), and red sea bream, Pagrus major (Temminck et Schlegel), revealed that the latter represent two new species, Philometra isaki sp. n. and Philometra madai sp. n., respectively. Molecular comparison of ITS2 rDNA between P. lateolabracis and P. madai supported the morphological conclusion that the two nematodes obtained from different fish species should be assigned to different species.